KR102392319B1 - 99m-Tc-labeled isonitrile-containing glucose derivative and method and use thereof - Google Patents

99m-Tc-labeled isonitrile-containing glucose derivative and method and use thereof Download PDF

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KR102392319B1
KR102392319B1 KR1020197028088A KR20197028088A KR102392319B1 KR 102392319 B1 KR102392319 B1 KR 102392319B1 KR 1020197028088 A KR1020197028088 A KR 1020197028088A KR 20197028088 A KR20197028088 A KR 20197028088A KR 102392319 B1 KR102392319 B1 KR 102392319B1
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cndg
appropriate amount
isonitrile
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KR20190121824A (en
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준보 장
슈란 장
취앤취앤 간
신 샤오
수에빈 왕
지강 탕
지에 루
잔빈 장
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베이징 시홍 파마수티컬 리서치 센터
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H13/00—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids
    • C07H13/02—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids
    • C07H13/04—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids having the esterifying carboxyl radicals attached to acyclic carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/0491—Sugars, nucleosides, nucleotides, oligonucleotides, nucleic acids, e.g. DNA, RNA, nucleic acid aptamers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives

Abstract

본 발명은 구조 일반식이 [99mTc-(CNDG)6]+인 99mTc 표지 이소니트릴-함유 글루코스 유도체 및 그 제조방법과 용도에 관한 것이다. 상기 유도체는 99mTc+을 중심핵으로 하고 CNDG 중 이소니트릴의 탄소원자가 99mTc(I)에 배위되어 6배위의 [99mTc-(CNDG)6]+ 배위화합물을 형성한다. [99mTc-(CNDG)6]+ 유도체는 리간드 CNDG의 합성 및 CNDG 동결건조 키트의 제조에 의하여 얻는다. 본 발명의 유도체는 안전성이 우수하고, 제조가 간편하며, 종양 부위에서 섭취가 높고 체류가 우수하며, 종양/비표적의 비율값이 높으며, 종양 영상에 사용 가능한 성능이 우수한 신규 99mTc 표지 이소니트릴-함유 글루코스 유도체로서, 보급화 및 응용에 유리하다.The present invention relates to a 99m Tc-labeled isonitrile-containing glucose derivative having the general formula [ 99m Tc-(CNDG) 6 ] + , and a method for preparing and use thereof. The derivative has 99m Tc + as the core and the carbon atom of isonitrile in CNDG is coordinated to 99m Tc(I) to form a 6-coordinated [ 99m Tc-(CNDG) 6 ] + coordination compound. [ 99m Tc-(CNDG) 6 ] + The derivative was obtained by synthesis of the ligand CNDG and preparation of a CNDG lyophilization kit. The derivative of the present invention is a novel 99m Tc-labeled isonitrile with excellent safety, simple preparation, high intake and excellent retention at the tumor site, high tumor/non-target ratio, and excellent performance for tumor imaging. -As a glucose derivative, it is advantageous for popularization and application.

Description

99m-Tc 표지 이소니트릴-함유 글루코스 유도체 및 그 제조방법과 용도99m-Tc-labeled isonitrile-containing glucose derivative and method and use thereof

본 발명은 방사성의약품화학 및 임상 핵의학 기술분야에 관한 것으로서, 구체적으로 99mTc 표지 이소니트릴-함유 글루코스 유도체 및 그 제조방법과 용도에 관한 것이다.The present invention relates to the technical field of radiopharmaceutical chemistry and clinical nuclear medicine, and more particularly, to a 99m Tc-labeled isonitrile-containing glucose derivative, and a method for preparing and use thereof.

현재, 임상의학 분야에서 악성 종양은 이미 인간의 건강과 생명을 위협하는 가장 치명적인 킬러가 되었고, 종양의 연간 발병률과 사망률은 여전히 상승하는 추세이며, 종양에 대한 조기 진단, 조기 치료 및 맞춤형 치료는 종양으로 인한 사망률을 감소시키는 가장 효과적인 방법이다. X선법, CT, MRI와 같은 기존의 비침습적인 종양 검출 영상 방법은 주로 병리적 상태에서의 장기의 형태와 구조의 영상이나, 양전자 방출 단층촬영(PET) 및 단일광자 방출 단층촬영(SPECT)과 같은 핵의학 분자영상 기술은 분자 수준에서 종양의 생리적, 병리적, 대사 및 기능의 변화를 반영할 수 있다. 현재, PET/CT, PET/MR 및 SPECT/CT 등 영상 기술의 유기적 융합이 종양의 조기 진단과 맞춤형 치료 면에서 더욱 중요한 역할을 발휘함에 따라, 임상 진단 및 치료에서의 핵의학 분자영상 기술의 광범위한 응용과 신속한 발전의 수요에 적응하기 위하여, 해당 기술이 의존하는 방사성 종양 분자 프로브에 대한 연구가 특히 시급하고 중요하다.At present, in the field of clinical medicine, malignant tumors have already become the most lethal killer of human health and life, and the annual incidence and mortality of tumors are still on the rise. It is the most effective way to reduce mortality due to Existing noninvasive tumor detection imaging methods such as X-ray, CT, and MRI are mainly images of the shape and structure of organs in pathological conditions, but positron emission tomography (PET) and single photon emission tomography (SPECT) and The same nuclear medicine molecular imaging technology can reflect changes in the physiology, pathology, metabolism and function of tumors at the molecular level. Currently, as the organic convergence of imaging technologies such as PET/CT, PET/MR and SPECT/CT plays a more important role in early diagnosis and customized treatment of tumors, a wide range of nuclear medicine molecular imaging technologies in clinical diagnosis and treatment has been developed. In order to adapt to the demands of applications and rapid development, the study of radioactive tumor molecular probes on which this technology depends is particularly urgent and important.

현재, 임상에서 가장 많이 사용되는 종양 영상제는 18F-플루오로데옥시글루코스(18F-FDG)이며, 영상 효과는 우수하나 고가의 진단 비용으로 인하여 광범위한 응용은 일정 정도의 제한을 받으며, 특히 경제적으로 뒤떨어진 국가와 지역에서의 응용은 아직 보급화 되지 않았다. 99mTc는 적절한 반감기(T1/2=6.02h)와 140 kev의 γ 단일광자 방출 등의 장점을 갖고 있고, 99Mo-99mTc 발생기의 보급화 및 응용으로 99mTc 방사성의약품을 간편하게 제조할 수 있고, 키트 형태로 제조 가능하며, 가격이 저렴하고 쉽게 이용 가능하며, 품질 신뢰성이 우수하기에, 임상 응용 가치가 있는 신규 99mTc 표지 글루코스계 종양 영상제의 개발은 아주 중요한 현실적 의의가 있다. 이소니트릴은 일반식이 RNC인 유기화합물이며, 그중 질소 원자는 부분 양전하를 띠고, 탄소 원자는 부분 음전하를 띤다. 연구에 따르면, 이소니트릴의 탄소 원자는 99mTc(I)에 배위되어 +1가의 [99mTc-(CNR)6]+ 배위화합물을 형성할 수 있으며, 그중 99mTc-메톡시이소부틸이소니트릴(99mTc-MIBI)은 심근 관류 영상제로 임상에서 이미 광범위하게 응용되고 있으며, 임상에서 해당 영상제가 일정한 종양 친화 작용도 있음을 발견하였다. 이소니트릴 분자는 본 발명의 종양 분자 프로브에서 99mTc와 당 분자를 연결시켜 이중기능 링커의 역할을 하여, 종양 친화의 당 대사 기능과 99mTc 추적 기능을 통합시킨다. 상술한 배경에 기초하여, 성능이 우수한 99mTc 표지 글루코스계 종양 분자 프로브를 개발 및 탐구하여, 글루코사민을 이소니트릴-함유 글루코스 유도체(CNDG라고 약칭함)로 전환시킨 후, 이소니트릴 리간드의 탄소 원자를 이용하여 99mTc에 배위시켜 얻은 안정적인 99mTc 표지 이소니트릴-함유 글루코스 유도체를 종양 영상제로 사용하는 것은 중요한 과학적 의의와 광범위한 응용 전망을 갖고 있으며, 본 분야가 직면한 중요한 과제이기도 하다.Currently, the most commonly used tumor imaging agent in clinical practice is 18 F-fluorodeoxyglucose ( 18 F-FDG), which has excellent imaging effects, but its wide application is limited to a certain extent due to high diagnostic costs, particularly economically The application in countries and regions lagging behind in terms of technology has not yet been popularized. 99m Tc has advantages such as an appropriate half-life (T 1/2 =6.02h) and γ single photon emission of 140 kev, and it is possible to easily manufacture 99m Tc radiopharmaceuticals with the popularization and application of the 99 Mo- 99m Tc generator. The development of a novel 99m Tc-labeled glucose-based tumor imaging agent with clinical application value is of great practical significance because it can be manufactured in the form of a kit, is inexpensive, is easily available, and has excellent quality reliability. Isonitrile is an organic compound with the general formula RNC, among which a nitrogen atom has a partial positive charge and a carbon atom has a partial negative charge. Studies have shown that the carbon atom of isonitrile can be coordinated to 99m Tc(I) to form a +1 valent [ 99m Tc-(CNR) 6 ] + coordination compound, of which 99m Tc-methoxyisobutylisonitrile ( 99m Tc-MIBI) has already been widely applied in clinical practice as a myocardial perfusion imaging agent, and it has been found that the imaging agent also has a certain tumor-affinity action in clinical practice. The isonitrile molecule serves as a bifunctional linker by linking 99m Tc and a sugar molecule in the tumor molecule probe of the present invention, thereby integrating the tumor-affinity sugar metabolism function and 99m Tc tracking function. Based on the background described above, 99m Tc with excellent performance A stable 99m Tc label obtained by developing and exploring a labeled glucose-based tumor molecular probe, converting glucosamine into an isonitrile-containing glucose derivative (abbreviated as CNDG), and then coordinating to 99m Tc using a carbon atom of an isonitrile ligand. The use of isonitrile-containing glucose derivatives as tumor imaging agents has important scientific significance and broad application prospects, and is also an important challenge facing this field.

본 발명은 안정성이 우수하고, 제조가 간편하며, 종양 영상제에 사용되는 99mTc 표지 이소니트릴-함유 글루코스 유도체를 제공함과 동시에 그 제조방법을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a 99m Tc-labeled isonitrile-containing glucose derivative used in a tumor imaging agent, which has excellent stability, is simple to manufacture, and at the same time provides a method for preparing the same.

상기 목적을 달성하기 위하여, 본 발명은 구조 일반식은 [99mTc-(CNDG)6]+이며, 하기 구조식을 갖는 99mTc 표지 이소니트릴-함유 글루코스 유도체를 제공한다:In order to achieve the above object, the present invention provides a 99m Tc-labeled isonitrile-containing glucose derivative having the structural formula [ 99m Tc-(CNDG) 6 ] + , and having the following structural formula:

Figure 112019098005433-pct00001
Figure 112019098005433-pct00001

상기 구조식에서 99mTc+핵을 중심핵으로 하고 CNDG 중 이소니트릴의 탄소원자가 99mTc(I)에 배위되어 6배위의 [99mTc-(CNDG)6]+ 배위화합물을 형성하며, n은 2 내지 5의 정수이다. In the above structural formula, 99m Tc + nucleus is the central core, and the carbon atom of isonitrile in CNDG is coordinated to 99m Tc(I) to form a 6-coordinated [ 99m Tc-(CNDG) 6 ] + coordination compound, where n is 2 to 5 is an integer of .

또한, 본 발명은 99mTc 표지 이소니트릴-함유 글루코스 유도체의 제조방법을 제공하며, 제조단계는 하기와 같다:In addition, the present invention provides a method for preparing a 99m Tc-labeled isonitrile-containing glucose derivative, the preparation steps being as follows:

a: 리간드 CNDG의 합성:a: Synthesis of ligand CNDG:

화합물(II)은 하기 구조식을 가지며:Compound (II) has the structural formula:

Figure 112019098005433-pct00002
Figure 112019098005433-pct00002

적당량의 글루코사민 염산염을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올을 첨가하여 용해시킨 후, 적당량의 NaOH을 첨가하여 실온에서 30min 동안 교반하여 반응시킨다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 적당량의 화합물(II)을 함유하는 메탄올 용액을 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 용매를 제거하고, 칼럼 크로마토그래피로 정제(디클로로메탄-메탄올)하여 리간드 CNDG를 얻는다.An appropriate amount of glucosamine hydrochloride is weighed into a 25 mL round bottom flask, dissolved by adding anhydrous methanol, an appropriate amount of NaOH is added, and the reaction is stirred at room temperature for 30 min. Then, the reaction flask is placed in an ice-water bath, a methanol solution containing an appropriate amount of compound (II) is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued in an ice-water bath for 3 h. After completion of the reaction, the solvent is distilled under reduced pressure. was removed and purified by column chromatography (dichloromethane-methanol) to obtain the ligand CNDG.

구체적인 합성 경로는 하기와 같다:Specific synthetic routes are as follows:

Figure 112019098005433-pct00003
Figure 112019098005433-pct00003

b: [99mTc-(CNDG)6]+ 배위화합물의 제조:b: [ 99m Tc-(CNDG) 6 ] + Preparation of coordination compound:

CNDG 동결건조 키트의 제조: CNDG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CNDG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비한다.Preparation of CNDG freeze-drying kit: CNDG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, The vial contains CNDG 1.0mg, SnCl 2 ·2H 2 O 0.03 mg and an appropriate amount of sodium citrate, and is prepared by freeze-drying.

갓 세척한 적당량의 Na99mTcO4을 CNDG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CNDG)6]+ 배위화합물을 얻을 수 있다.An appropriate amount of freshly washed Na 99m TcO 4 After adding to the CNDG freeze-drying kit, shaking sufficiently, and after the solid is completely dissolved, the [ 99m Tc-(CNDG) 6 ] + coordination compound can be obtained by heating in a boiling water bath for 20 min.

구체적인 제조단계는 하기와 같다:The specific manufacturing steps are as follows:

1. CN2DG의 합성1. Synthesis of CN2DG

글루코사민 염산염 91mg(0.423mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 17mg(0.423mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=2) 114mg(0.466mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 CN2DG을 얻었다.91 mg (0.423 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, and then 17 mg (0.423 mmol) of NaOH was added thereto, followed by stirring at room temperature for 30 min. Then, the reaction flask is placed in an ice-water bath, and 1 mL of a methanol solution in which 114 mg (0.466 mmol) of compound II (n=2) is dissolved is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and dried to obtain CN2DG.

2. CN3DG의 합성2. Synthesis of CN3DG

글루코사민 염산염 117mg(0.544mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 22mg(0.544mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=3) 156mg(0.598mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 CN3DG을 얻었다.117 mg (0.544 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, and then 22 mg (0.544 mmol) of NaOH was added thereto, followed by stirring at room temperature for 30 min. Then, the reaction flask is placed in an ice-water bath, and 1 mL of a methanol solution in which 156 mg (0.598 mmol) of compound II (n=3) is dissolved is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and after drying, CN3DG was obtained.

3. CN4DG의 합성3. Synthesis of CN4DG

글루코사민 염산염 129mg(0.600mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 24mg(0.600mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=4) 182mg(0.660mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 CN4DG을 얻었다.129 mg (0.600 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, 24 mg (0.600 mmol) of NaOH was added thereto, and the reaction was stirred at room temperature for 30 min. Then, the reaction flask was placed in an ice-water bath, and 1 mL of a methanol solution in which 182 mg (0.660 mmol) of compound II (n = 4) was dissolved was slowly added dropwise under stirring, and after the dropping was completed, the reaction was continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and after drying, CN4DG was obtained.

4. CN5DG의 합성4. Synthesis of CN5DG

글루코사민 염산염 147mg(0.684mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 27mg(0.684mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=5) 217mg(0.752mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 CN5DG를 얻었다.147 mg (0.684 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, and then 27 mg (0.684 mmol) of NaOH was added, followed by stirring at room temperature for 30 min. Then, the reaction flask was placed in an ice-water bath, and 1 mL of a methanol solution in which 217 mg (0.752 mmol) of compound II (n=5) was dissolved was slowly added dropwise under stirring, and after the dropping was completed, the reaction was continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and dried to obtain CN5DG.

5. [99mTc-(CN2DG)6]+, [99mTc-(CN3DG)6]+, [99mTc-(CN4DG)6]+ 및 [99mTc-(CN5DG)6]+의 합성5. Synthesis of [ 99m Tc-(CN2DG) 6 ] + , [ 99m Tc-(CN3DG) 6 ] + , [ 99m Tc-(CN4DG) 6 ] + and [ 99m Tc-(CN5DG) 6 ] +

CN2DG 동결건조 키트의 제조: CN2DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN2DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN2DG freeze-drying kit: CN2DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, and each The vial contained 1.0 mg of CN2DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN2DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN2DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN2DG lyophilization kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN2DG) 6 ] + coordination compound can get

CN3DG 동결건조 키트의 제조: CN3DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN3DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN3DG freeze-drying kit: CN3DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, and each The vial contained 1.0 mg of CN3DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN3DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN3DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN3DG lyophilization kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN3DG) 6 ] + coordination compound can get

CN4DG 동결건조 키트의 제조: CN4DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN4DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN4DG lyophilization kit: CN4DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, The vial contained 1.0 mg of CN4DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN4DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN4DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN4DG freeze-drying kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN4DG) 6 ] + coordination compound can get

CN5DG 동결건조 키트의 제조: CN5DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN5DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN5DG freeze-drying kit: CN5DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, and each The vial contained 1.0 mg of CN5DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN5DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN5DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN5DG lyophilization kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN5DG) 6 ] + coordination compound can get

본 발명의 화학 합성에 사용되는 시약 원료는 모두 시판 제품이며, 광범위한 출처를 갖는다. 상기 방법으로 제조한 [99mTc-(CNDG)6]+ 배위화합물의 방사화학적 순도는 90%를 초과하고, 친수성 물질로서 체외 안전성이 우수하다. 상기 배위화합물은 종양 보유 마우스의 종양 부위에서 섭취가 높고 체류가 우수하며, 종양/근육 및 종양/혈액의 비율값이 우수하며, 간, 신장, 폐 등 비표적 장기에서의 섭취 수치가 낮은 바, 종양 영상에 사용 가능한 성능이 우수한 신규 99mTc 표지 이소니트릴-함유 글루코스 유도체이기에, 본 발명의 목적을 달성하였다.All reagent raw materials used in the chemical synthesis of the present invention are commercially available products and have a wide range of sources. The radiochemical purity of the [ 99m Tc-(CNDG) 6 ] + coordination compound prepared by the above method exceeds 90%, and as a hydrophilic material, it has excellent in vitro safety. The coordination compound has high intake and excellent retention at the tumor site of tumor-bearing mice, excellent tumor/muscle and tumor/blood ratio values, and low intake levels in non-target organs such as liver, kidney, and lung, Since it is a novel 99m Tc-labeled isonitrile-containing glucose derivative with excellent performance usable for tumor imaging, the object of the present invention was achieved.

본 발명의 [99mTc-(CNDG)6]+의 성능 측정:Performance measurement of [ 99m Tc-(CNDG) 6 ] + of the present invention:

1. [99mTc-(CNDG)6]+ 배위화합물의 크로마토그래피 검증1. [ 99m Tc-(CNDG) 6 ] + Chromatographic verification of coordination compound

박층 크로마토그래피법(TLC)으로 표지물의 방사화학적 순도를 검증하며, 사용된 전개 시스템은 메탄올/폴리아미드 박막 시트이며, 해당 시스템에서 각 방사성 성분의 Rf값은 하기 표에 나타낸 바와 같다.The radiochemical purity of the label was verified by thin layer chromatography (TLC), and the developing system used was a methanol/polyamide thin film sheet, and the R f values of each radioactive component in the system are as shown in the table below.

Figure 112019098005433-pct00004
Figure 112019098005433-pct00004

상기 크로마토그래피 검증에 의해 측정된 표지물의 방사화학적 순도는 모두 90%를 초과한다.The radiochemical purity of the labels as determined by the chromatographic verification above all exceeded 90%.

2. 지질-물 분배계수의 측정2. Determination of the lipid-water partition coefficient

생리식염수로 희석한 표지용액 100μL(50μCi)를 취하여 2mL 원심분리관에 투입한 후, 원심분리관에 n-옥탄올 800μL와 PBS 700μL를 첨가하고, 원심분리관 덮개를 덮어서 3min(2500r/min) 동안 볼텍싱하며, 용액이 층분리가 될 때까지 정치한 후 원심분리기에서 3min(3000r/min) 동안 원심분리하고, 상부 유기상과 하부 수상을 각각 100μL씩 3부를 취하여, γ-counter에서 유기상과 수상의 방사계수를 측정하였다. 지질-물 분배계수 P = 유기상 방사계수/수상 방사계수이며, 일반적으로 지질-물 분배계수는 Log P로 표시한다. 측정을 통하여, [99mTc-(CN2DG)6]+, [99mTc-(CN3DG)6]+, [99mTc-(CN4DG)6]+ 및 [99mTc-(CN5DG)6]+의 Log P값은 각각 -4.26±0.12, -4.01±0.17, -3.84±0.06, -3.57±0.35이며, 이는 표지물이 모두 수용성 물질이고, n값이 증가함에 따라 해당 표지물의 친수 성능이 감소됨을 설명한다.Take 100 μL (50 μCi) of the labeling solution diluted with physiological saline, put it in a 2 mL centrifuge tube, add 800 μL of n-octanol and 700 μL of PBS to the centrifuge tube, and cover the centrifuge tube for 3 min (2500 r/min) After vortexing for a while, the solution was allowed to stand until the layers were separated, centrifuged for 3 min (3000 r/min) in a centrifuge, and 3 parts of the upper organic phase and the lower aqueous phase were taken 100 μL each, and the organic phase and the aqueous phase in the γ-counter of was measured. The lipid-water partition coefficient P = organic phase radiation coefficient/water phase radiation coefficient, and the lipid-water partition coefficient is generally expressed as Log P. Through the measurements, [ 99m Tc-(CN2DG) 6 ] + , [ 99m Tc-(CN3DG) 6 ] + , [ 99m Tc-(CN4DG) 6 ] + and [ 99m Tc-(CN5DG) 6 ] + Log P values are -4.26±0.12, -4.01±0.17, -3.84±0.06, and -3.57±0.35, respectively, which explains that all of the labels are water-soluble substances, and that the hydrophilic performance of the label decreases as the n value increases. .

3. 안정성의 측정3. Measurement of Stability

표지물을 각각 실온 및 37℃에서 마우스의 혈청 내에 시간별(1, 2, 3, 4시간)로 방치한 후 방사화학적 순도를 측정하였다. 실험 결과, 각 표지물이 실온 및 37℃에서 마우스의 혈청 내에 4시간 동안 방치된 후의 방사화학적 순도는 모두 90%를 초과하며, 이는 표지물의 체외 안정성이 우수함을 설명한다.After the label was left in the serum of mice at room temperature and 37° C. for each hour (1, 2, 3, 4 hours), the radiochemical purity was measured. As a result of the experiment, the radiochemical purity of each label after leaving it in the serum of mice at room temperature and 37°C for 4 hours all exceeded 90%, which explains the excellent in vitro stability of the label.

4. 종양 보유 마우스 체내에서의 생체분포의 측정4. Measurement of biodistribution in tumor-bearing mice

표지용액(0.1mL, 74KBq)을 꼬리 정맥을 통하여 마우스 체내에 주입하고, 주사 시간을 기록한 후, 각 시점(30min, 60min, 120min)에서 마우스를 경추 탈구하여 희생시키고(각 시점 5마리 마우스), 해부 후 심장, 간, 폐, 신장, 비장, 위, 근육, 혈액, 종양 등 관심있는 장기를 취하여, γ-counter로 각 장기의 방사계수를 측정하고, 각 장기의 질량을 기준으로 환산하여 각 장기의 섭취 수치(단위 %ID/g)를 얻었으며, 각 표지물의 종양 보유 마우스 체내에서의 생체분포 결과를 표1 내지 표4에 나타내었다.After injecting the labeling solution (0.1mL, 74KBq) into the mouse body through the tail vein, recording the injection time, the mice were sacrificed by cervical dislocation at each time point (30min, 60min, 120min) (5 mice at each time point), After dissection, organs of interest such as heart, liver, lung, kidney, spleen, stomach, muscle, blood, and tumor are taken, and the radiation coefficient of each organ is measured with a γ-counter, and each organ is converted based on the mass of each organ. was obtained, and the biodistribution results of each marker in tumor-bearing mice are shown in Tables 1 to 4.

Figure 112019098005433-pct00005
Figure 112019098005433-pct00005

Figure 112019098005433-pct00006
Figure 112019098005433-pct00006

Figure 112019098005433-pct00007
Figure 112019098005433-pct00007

Figure 112019098005433-pct00008
Figure 112019098005433-pct00008

[99mTc-(CN2DG)6]+, [99mTc-(CN3DG)6]+, [99mTc-(CN4DG)6]+ 및 [99mTc-(CN5DG)6]+과 이미 제3상 임상 연구에 진입한 99mTc-ECDG(David J. et al Imaging with 99mTc-ECDG Targeted at the Multifunctional Glucose Transport System: Feasibility Study with Rodents [J]. Radiology, 2003,226(2):465-473)의 종양 보유 마우스 체내에서의 생체분포 데이터를 비교하였으며, 그 결과를 표5에 나타내었다.[ 99m Tc-(CN2DG) 6 ] + , [ 99m Tc-(CN3DG) 6 ] + , [ 99m Tc-(CN4DG) 6 ] + and [ 99m Tc-(CN5DG) 6 ] + and already in phase 3 clinical studies 99m Tc-ECDG (David J. et al Imaging with 99m Tc-ECDG Targeted at the Multifunctional Glucose Transport System: Feasibility Study with Rodents [J]. Radiology, 2003,226(2):465-473) The biodistribution data in the body of the mice were compared, and the results are shown in Table 5.

Figure 112019098005433-pct00009
Figure 112019098005433-pct00009

상기 결과에 의하면, [99mTc-(CN2DG)6]+, [99mTc-(CN3DG)6]+, [99mTc-(CN4DG)6]+ 및 [99mTc-(CN5DG)6]+는 종양에서의 섭취, 종양/혈액 및 종양/간 비율값이 모두 99mTc-ECDG보다 우수하고, 간, 신장 등 장기에서의 섭취는 모두 99mTc-ECDG보다 낮기에, 성능이 우수한 신규 종양 영상제로 널리 응용될 수 있다.According to the above results, [ 99m Tc-(CN2DG) 6 ] + , [ 99m Tc-(CN3DG) 6 ] + , [ 99m Tc-(CN4DG) 6 ] + and [ 99m Tc-(CN5DG) 6 ] + is superior to 99m Tc-ECDG in uptake from tumor, tumor/blood, and tumor/liver ratio, and uptake from organs such as liver and kidney is all higher than 99m Tc -ECDG Because it is low, it can be widely applied as a novel tumor imaging agent with excellent performance.

5. SPECT 영상의 측정5. SPECT IMAGE MEASUREMENT

제조된 [99mTc-(CN3DG)6]+ 또는 [99mTc-(CN5DG)6]+(0.2mL, 700μCi)을 꼬리 정맥을 통하여 종양 보유 마우스 체내에 주입하고, 1h후에 SPECT 영상을 수행하였다. SPECT 영상 결과, 양자는 모두 종양 부위에 확연히 집중되었고, 신장에서도 비교적 높은 농도를 가지나, 다른 장기에서의 섭취가 낮기에, 성능이 우수한 종양 영상제로 될 수 있음을 나타낸다.prepared [ 99m Tc-(CN3DG) 6 ] + or [ 99m Tc-(CN5DG) 6 ] + (0.2mL, 700μCi) was injected into the tumor-bearing mouse body through the tail vein, and SPECT imaging was performed after 1 h. As a result of SPECT imaging, both were clearly concentrated in the tumor site, and although they had a relatively high concentration in the kidney, their intake in other organs was low, indicating that they could be excellent tumor imaging agents.

이하, 실시예를 통하여 본 발명을 상세하게 설명한다. 99mTc 표지 이소니트릴-함유 글루코스 유도체의 구조 일반식은 [99mTc-(CNDG)6]+이며, 제조단계는 하기와 같다:Hereinafter, the present invention will be described in detail through examples. The structural formula of the 99m Tc-labeled isonitrile-containing glucose derivative is [ 99m Tc-(CNDG) 6 ] + , and the preparation steps are as follows:

a: 리간드 CNDG의 합성:a: Synthesis of ligand CNDG:

적당량의 글루코사민 염산염을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올을 첨가하여 용해시킨 후, 적당량의 NaOH을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 적당량의 화합물(II)을 함유하는 메탄올 용액을 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 용매를 제거하고, 칼럼 크로마토그래피로 정제(디클로로메탄-메탄올)하여 리간드 CNDG를 얻었다.An appropriate amount of glucosamine hydrochloride was weighed into a 25 mL round bottom flask, dissolved by adding anhydrous methanol, followed by addition of an appropriate amount of NaOH, followed by stirring at room temperature for 30 min. Then, the reaction flask is placed in an ice-water bath, a methanol solution containing an appropriate amount of compound (II) is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued in an ice-water bath for 3 h. After completion of the reaction, the solvent is distilled under reduced pressure. was removed and purified by column chromatography (dichloromethane-methanol) to obtain a ligand CNDG.

구체적인 합성 경로는 하기와 같다:Specific synthetic routes are as follows:

Figure 112019098005433-pct00010
Figure 112019098005433-pct00010

b: [99mTc-(CNDG)6]+ 배위화합물의 제조, 구체적인 제조단계는 하기와 같다:b: [ 99m Tc-(CNDG) 6 ] + Preparation of coordination compound, specific preparation steps are as follows:

1. CN2DG의 합성1. Synthesis of CN2DG

글루코사민 염산염 91mg(0.423mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 17mg(0.423mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=2) 114mg(0.466mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 생성물 CN2DG 83mg을 얻었으며, 수율은 75%이다. 1H-NMR(400 MHz, D2O): δ (ppm) 3.65-3.89 (m, 7H), 3.34-3.44 (m, 2H), 2.65 (t, 2H). HRMS Calculated for, C10H16N2O6Na [M+Na]+ 283.0911, found 283.0906. IR (KBr)/ cm-1: 3303.24 (-OH), 2933.85, 2149.76 (-NC), 1647.28 (-C=O), 1560.48, 1303.03, 586.39.91 mg (0.423 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, and then 17 mg (0.423 mmol) of NaOH was added thereto, followed by stirring at room temperature for 30 min. Then, the reaction flask is placed in an ice-water bath, and 1 mL of a methanol solution in which 114 mg (0.466 mmol) of compound II (n=2) is dissolved is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and after drying, 83mg of the product CN2DG was obtained, and the yield was 75%. 1 H-NMR (400 MHz, D 2 O): δ (ppm) 3.65-3.89 (m, 7H), 3.34-3.44 (m, 2H), 2.65 (t, 2H). HRMS Calculated for, C 10 H 16 N 2 O 6 Na [M+Na] + 283.0911, found 283.0906. IR (KBr) / cm -1 : 3303.24 (-OH), 2933.85, 2149.76 (-NC), 1647.28 (-C=O), 1560.48, 1303.03, 586.39.

2. CN3DG의 합성2. Synthesis of CN3DG

글루코사민 염산염 117mg(0.544mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 22mg(0.544mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=3) 156mg(0.598mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 생성물 CN3DG 85mg을 얻었으며, 수율은 57%이다. 1H-NMR(400 MHz, D2O): δ (ppm) 3.65-3.85 (m, 5H), 3.42-3.49 (m, 4H), 2.37-2.42 (m, 2H), 2.19 (t, 2H). HRMS Calculated for C11H18N2O6Na [M+Na]+, 297.1057, found, 297.1052. IR (KBr)/ cm-1: 3299.38 (-OH), 2932.89, 2149.76 (-NC), 1647.28 (-C=O), 1558.55,1303.03, 588.31.117 mg (0.544 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, and then 22 mg (0.544 mmol) of NaOH was added thereto, followed by stirring at room temperature for 30 min. Then, the reaction flask is placed in an ice-water bath, and 1 mL of a methanol solution in which 156 mg (0.598 mmol) of compound II (n=3) is dissolved is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and after drying, 85 mg of the product CN3DG was obtained, and the yield was 57%. 1 H-NMR (400 MHz, D 2 O): δ (ppm) 3.65-3.85 (m, 5H), 3.42-3.49 (m, 4H), 2.37-2.42 (m, 2H), 2.19 (t, 2H) . HRMS Calculated for C 11 H 18 N 2 O 6 Na [M+Na] + , 297.1057, found, 297.1052. IR (KBr)/cm -1 : 3299.38 (-OH), 2932.89, 2149.76 (-NC), 1647.28 (-C=O), 1558.55,1303.03, 588.31.

3. CN4DG의 합성3. Synthesis of CN4DG

글루코사민 염산염 129mg(0.600mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 24mg(0.600mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=4) 182mg(0.660mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 생성물 CN4DG 108mg을 얻었으며, 수율은 63%이다. 1H-NMR(400 MHz, D2O): δ (ppm) 3.65-3.89 (m, 5H), 3.38-3.44 (m, 4H), 2.28 (t, 2H), 1.60-1.75 (m, 4H). HRMS Calculated for C12H21N2O6 [M+H]+, 289.1394, found 289.1396. IR (KBr)/ cm-1: 3295.52 (-OH), 2943.50, 2150.72 (-NC), 1645.35 (-C=O), 1542.15, 1093.69, 1033.89, 599.89.129 mg (0.600 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, 24 mg (0.600 mmol) of NaOH was added thereto, and the reaction was stirred at room temperature for 30 min. Then, the reaction flask was placed in an ice-water bath, and 1 mL of a methanol solution in which 182 mg (0.660 mmol) of compound II (n = 4) was dissolved was slowly added dropwise under stirring, and after the dropping was completed, the reaction was continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and after drying, 108mg of the product CN4DG was obtained, and the yield was 63%. 1 H-NMR (400 MHz, D 2 O): δ (ppm) 3.65-3.89 (m, 5H), 3.38-3.44 (m, 4H), 2.28 (t, 2H), 1.60-1.75 (m, 4H) . HRMS Calculated for C 12 H 21 N 2 O 6 [M+H] + , 289.1394, found 289.1396. IR (KBr)/cm -1 : 3295.52 (-OH), 2943.50, 2150.72 (-NC), 1645.35 (-C=O), 1542.15, 1093.69, 1033.89, 599.89.

4. CN5DG의 합성4. Synthesis of CN5DG

글루코사민 염산염 147mg(0.684mmol)을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올 3mL를 첨가하여 용해시킨 후, NaOH 27mg(0.684mmol)을 첨가하여 실온에서 30min 동안 교반하여 반응시켰다. 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 화합물II(n=5) 217mg(0.752mmol)이 용해된 메탄올 용액 1mL를 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 메탄올 용매를 제거하고, 잔류 고형물을 실리카겔 칼럼(디클로로메탄:메탄올=5:1)으로 분리하여 정제하며, 건조 후 생성물 CN5DG 161mg를 얻었으며, 수율은 78%이다. 1H-NMR(400 MHz, D2O): δ (ppm) 3.74-3.82 (m, 3H), 3.63-3.70 (m, 2H), 3.34-3.40 (m, 4H), 2.24 (q, 2H),1.53-1.59 (m, 4H), 1.33-1.38 (m, 2H). HRMS Calculated for C13H22N2O6Na [M+Na]+, 325.1374, found 325.1370. IR (KBr)/ cm-1: 3294.56 (-OH), 2941.57, 2148.79 (-NC), 1645.35 (-C=O), 1538.30, 1093.69, 1032.93, 590.85.147 mg (0.684 mmol) of glucosamine hydrochloride was weighed in a 25 mL round bottom flask, dissolved by adding 3 mL of anhydrous methanol, and then 27 mg (0.684 mmol) of NaOH was added, followed by stirring at room temperature for 30 min. Then, the reaction flask was placed in an ice-water bath, and 1 mL of a methanol solution in which 217 mg (0.752 mmol) of compound II (n=5) was dissolved was slowly added dropwise under stirring, and after the dropping was completed, the reaction was continued for 3 h in an ice-water bath, After completion of the reaction, the methanol solvent was removed by distillation under reduced pressure, and the residual solid was separated and purified by a silica gel column (dichloromethane:methanol=5:1), and after drying, 161mg of the product CN5DG was obtained, and the yield was 78%. 1 H-NMR (400 MHz, D 2 O): δ (ppm) 3.74-3.82 (m, 3H), 3.63-3.70 (m, 2H), 3.34-3.40 (m, 4H), 2.24 (q, 2H) , 1.53-1.59 (m, 4H), 1.33-1.38 (m, 2H). HRMS Calculated for C 13 H 22 N 2 O 6 Na [M+Na] + , 325.1374, found 325.1370. IR (KBr)/cm -1 : 3294.56 (-OH), 2941.57, 2148.79 (-NC), 1645.35 (-C=O), 1538.30, 1093.69, 1032.93, 590.85.

5. [99mTc-(CN2DG)6]+, [99mTc-(CN3DG)6]+, [99mTc-(CN4DG)6]+ 및 [99mTc-(CN5DG)6]+의 합성5. Synthesis of [ 99m Tc-(CN2DG) 6 ] + , [ 99m Tc-(CN3DG) 6 ] + , [ 99m Tc-(CN4DG) 6 ] + and [ 99m Tc-(CN5DG) 6 ] +

CN2DG 동결건조 키트의 제조: CN2DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN2DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN2DG freeze-drying kit: CN2DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, and each The vial contained 1.0 mg of CN2DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN2DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN2DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN2DG lyophilization kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN2DG) 6 ] + coordination compound can get

CN3DG 동결건조 키트의 제조: CN3DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN3DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN3DG freeze-drying kit: CN3DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, and each The vial contained 1.0 mg of CN3DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN3DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN3DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN3DG lyophilization kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN3DG) 6 ] + coordination compound can get

CN4DG 동결건조 키트의 제조: CN4DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN4DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN4DG freeze-drying kit: CN4DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is placed in a clean penicillin vial, The vial contained 1.0 mg of CN4DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN4DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN4DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN4DG freeze-drying kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN4DG) 6 ] + coordination compound can get

CN5DG 동결건조 키트의 제조: CN5DG, SnCl2·2H2O, 구연산나트륨 등을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CN5DG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하였다.Preparation of CN5DG freeze-drying kit: CN5DG, SnCl 2 .2H 2 O, sodium citrate, etc. are dissolved in an appropriate amount of re-distilled water, the pH is adjusted to 5 to 6, and after sufficient dissolution, 1 mL each is put into a clean penicillin vial, and each The vial contained 1.0 mg of CN5DG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, and was prepared by freeze-drying.

갓 세척한 Na99mTcO4 1 내지 5mL를 CN5DG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CN5DG)6]+ 배위화합물을 얻을 수 있다.1 to 5 mL of freshly washed Na 99m TcO 4 is added to the CN5DG lyophilization kit, shaken sufficiently, and after the solid is completely dissolved, heated in a boiling water bath for 20 min. [ 99m Tc-(CN5DG) 6 ] + coordination compound can get

상기 99mTc 표지 이소니트릴-함유 글루코스 유도체는 종양 영상제로서 핵의학 분야에 응용될 수 있다.The 99m Tc-labeled isonitrile-containing glucose derivative can be applied to nuclear medicine as a tumor imaging agent.

상술한 실시예는 본 발명의 내용을 설명하기 위한 것일 뿐, 본 발명의 범위를 한정하기 위한 것이 아니다.The above-described embodiment is only for explaining the content of the present invention, not for limiting the scope of the present invention.

본 발명은 구조 일반식이 [99mTc-(CNDG)6]+인 99mTc 표지 이소니트릴-함유 글루코스 유도체 및 그 제조방법과 용도를 제공한다. 상기 유도체는 99mTc+을 중심핵으로 하고 CNDG 중 이소니트릴의 탄소원자가 99mTc(I)에 배위되어 6배위의 [99mTc-(CNDG)6]+ 배위화합물을 형성한다. 본 발명에서 제공하는 유도체는 안전성이 우수하고, 제조가 간편하며, 종양부위에서 섭취가 높고 체류가 우수하며, 종양/비표적의 비율값이 높으며, 종양 영상에 사용 가능한 성능이 우수한 신규 99mTc 표지 이소니트릴-함유 글루코스 유도체로서, 보급화 및 응용에 유리하며, 경제적 가치와 응용 가능성이 높다.The present invention provides a 99m Tc-labeled isonitrile-containing glucose derivative having the general formula [ 99m Tc-(CNDG) 6 ] + , and a preparation method and use thereof. The derivative has 99m Tc + as the core and the carbon atom of isonitrile in CNDG is coordinated to 99m Tc(I) to form a 6-coordinated [ 99m Tc-(CNDG) 6 ] + coordination compound. The derivative provided in the present invention is a novel 99m Tc marker with excellent safety, simple preparation, high intake and excellent retention at the tumor site, high tumor/non-target ratio, and excellent performance for tumor imaging. As an isonitrile-containing glucose derivative, it is advantageous for popularization and application, and has high economic value and application potential.

Claims (3)

구조 일반식은 [99mTc-(CNDG)6]+이며, 하기 구조식을 갖는 99mTc 표지 이소니트릴-함유 글루코스 유도체:
Figure 112021141228207-pct00011

상기 구조식에서 99mTc+핵을 중심핵으로 하고 CNDG 중 이소니트릴의 탄소원자가 99mTc(I)에 배위되어 6배위의 [99mTc-(CNDG)6]+ 배위화합물을 형성하며, n은 2 내지 5의 정수이다.The general formula is [ 99m Tc-(CNDG) 6 ] + , and a 99m Tc labeled isonitrile-containing glucose derivative having the structural formula:
Figure 112021141228207-pct00011

In the above structural formula, 99m Tc + nucleus is the central core, and the carbon atom of isonitrile in CNDG is coordinated to 99m Tc(I) to form a 6-coordinated [ 99m Tc-(CNDG) 6 ] + coordination compound, where n is 2 to 5 is the integer of a: 리간드 CNDG의 합성:
화합물(II)은 하기 구조식을 가지며:
Figure 112021065459567-pct00012

적당량의 글루코사민 염산염을 25mL 둥근 바닥 플라스크에 칭량하고, 무수 메탄올을 첨가하여 용해시킨 후, 적당량의 NaOH을 첨가하여 실온에서 30min 동안 교반하여 반응시키며; 그 다음, 반응 플라스크를 빙수욕에 넣고, 교반하에 적당량의 화합물(II)을 함유하는 메탄올 용액을 서서히 적하하며, 적하가 완료된 후 계속하여 빙수욕에서 3h 동안 반응시키며, 반응 종료 후 감압 증류하여 용매를 제거하고, 칼럼 크로마토그래피로 정제하여 리간드 CNDG를 얻으며,
구체적인 합성 경로는 하기와 같으며:
Figure 112021065459567-pct00013

b: [99mTc-(CNDG)6]+ 배위화합물의 제조:
CNDG 동결건조 키트의 제조: CNDG, SnCl2·2H2O, 구연산나트륨을 적당량의 재증류수에 용해시키고, pH를 5 내지 6으로 조정하며, 충분히 용해시킨 후 깨끗한 페니실린 바이알에 1mL씩 넣고, 각 바이알은 CNDG 1.0mg, SnCl2·2H2O 0.03mg 및 적당량의 구연산나트륨을 함유하며, 동결건조하여 준비하며;
갓 세척한 적당량의 Na99mTcO4을 CNDG 동결건조 키트에 첨가하고, 충분히 흔들어 주며, 고형물이 완전히 용해된 후, 비등 수욕에서 20min 동안 가열하면 상기 [99mTc-(CNDG)6]+ 배위화합물을 얻을 수 있는; 제조단계를 포함하는 제1항의 99mTc 표지 이소니트릴-함유 글루코스 유도체의 제조방법.a: Synthesis of ligand CNDG:
Compound (II) has the structural formula:
Figure 112021065459567-pct00012

An appropriate amount of glucosamine hydrochloride was weighed into a 25 mL round bottom flask, dissolved by adding anhydrous methanol, followed by addition of an appropriate amount of NaOH and stirred at room temperature for 30 min to react; Then, the reaction flask is placed in an ice-water bath, a methanol solution containing an appropriate amount of compound (II) is slowly added dropwise under stirring, and after completion of the dropping, the reaction is continued in an ice-water bath for 3 h. After completion of the reaction, the solvent is distilled under reduced pressure. Removed, purified by column chromatography to obtain ligand CNDG,
Specific synthetic routes are as follows:
Figure 112021065459567-pct00013

b: [ 99m Tc-(CNDG) 6 ] + Preparation of coordination compound:
Preparation of CNDG freeze-drying kit: Dissolve CNDG, SnCl 2 .2H 2 O, and sodium citrate in an appropriate amount of re-distilled water, adjust the pH to 5 to 6, and after sufficient dissolution, put 1 mL into a clean penicillin vial, each vial Silver contains 1.0 mg of CNDG, 0.03 mg of SnCl 2 ·2H 2 O and an appropriate amount of sodium citrate, prepared by freeze-drying;
An appropriate amount of freshly washed Na 99m TcO 4 After adding to the CNDG freeze-drying kit, shaking sufficiently, and after completely dissolving the solid, heating in a boiling water bath for 20 min to obtain the [ 99m Tc-(CNDG) 6 ] + coordination compound; A method for preparing the 99m Tc-labeled isonitrile-containing glucose derivative of claim 1, comprising the preparation step. 삭제delete
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